IL1F10 Human His

What is IL1F10 and what are its common alternative names in scientific literature?

IL1F10 (Interleukin 1 Family Member 10) is a protein-coding gene that produces a cytokine belonging to the interleukin-1 family. The protein is commonly referenced under multiple names in scientific literature, including IL-38, IL-1F10, IL-1 Theta, IL-1HY2, FIL1 Theta, and FKSG75 . When designing literature searches or comparing research findings, it's essential to incorporate all these alternative nomenclatures to ensure comprehensive coverage. The current consensus nomenclature favors IL-38 in more recent publications, though many databases and repositories still primarily list the protein under IL1F10 .

What functional roles does IL1F10/IL-38 play in immune regulation?

IL1F10/IL-38 functions as an immunomodulatory cytokine with primarily anti-inflammatory properties in specific contexts. Research indicates that the protein itself does not directly induce cytokine production but rather modulates existing inflammatory pathways . It has been shown to reduce IL22 and IL17A production by T-cells responding to heat-killed Candida albicans and decreases IL36G-induced production of IL8 by peripheral blood mononuclear cells . Conversely, it can increase IL6 production by dendritic cells stimulated with bacterial lipopolysaccharides (LPS) . This context-dependent immunomodulation makes IL1F10 an interesting target for studying complex immune regulation. Methodologically, researchers should consider measuring multiple cytokine outputs simultaneously when examining IL1F10 activity rather than focusing on single readouts.

What expression systems are commonly used for producing His-tagged IL1F10, and how do they differ?

Recombinant His-tagged IL1F10 protein can be produced in various expression systems, with E. coli and yeast being the most commonly documented . The choice of expression system significantly impacts protein characteristics:

• E. coli expression: Provides high yield and is cost-effective but may result in proteins lacking proper folding or post-translational modifications. The recombinant proteins produced in E. coli systems typically have purity >98% as determined by reducing SDS-PAGE .

• Yeast expression: Offers mammalian-like post-translational modifications while maintaining relatively high yield. Yeast-expressed IL1F10 His-tagged proteins typically achieve >90% purity as determined by SDS-PAGE .

When designing experiments, researchers should consider whether post-translational modifications are critical for their specific research questions. For structural studies, E. coli-derived protein may be sufficient, while functional studies examining receptor interactions might benefit from yeast or mammalian cell-expressed proteins.

What are the optimal reconstitution and storage conditions for lyophilized His-tagged IL1F10?

Lyophilized His-tagged IL1F10 recombinant protein requires careful handling to maintain activity. The recommended reconstitution protocol involves dissolving the lyophilized protein in sterile PBS (pH 7.4) . Once reconstituted, the protein solution exhibits different stability profiles depending on storage temperature:

• Short-term storage (2-7 days): 4-8°C is sufficient

• Medium-term storage (up to 3 months): Aliquot and store at ≤-20°C

• Long-term storage (up to 12 months): Store at -20°C to -80°C

To maintain protein integrity, researchers should avoid repeated freeze-thaw cycles by preparing single-use aliquots immediately after reconstitution. Additionally, it's advisable to add carrier protein (such as 0.1% BSA) when diluting to working concentrations for increased stability during experimental procedures.

How can researchers validate the purity and activity of His-tagged IL1F10 preparations?

Validation of His-tagged IL1F10 involves multiple complementary techniques:

• Purity assessment: SDS-PAGE under reducing conditions is the standard method, with commercial preparations typically showing >90-98% purity . Western blotting using anti-His antibodies provides additional confirmation of identity.

• Functional validation: Unlike many cytokines that directly induce responses, IL1F10 primarily modulates existing inflammatory pathways. Functional validation therefore requires:

  • Testing inhibition of IL22 and IL17A production by T-cells responding to heat-killed Candida albicans

  • Measuring reduction in IL36G-induced IL8 production by PBMCs

  • Assessing enhancement of LPS-induced IL6 production by dendritic cells

• Binding validation: Surface plasmon resonance or co-immunoprecipitation assays to confirm binding to IL-36R/IL1RL2

What cell types and experimental models are most appropriate for studying IL1F10 functions?

The selection of appropriate cellular and experimental models is crucial for investigating IL1F10 functions:

Cellular models:

• T-cells: For studying IL1F10's effects on cytokine production (IL22, IL17A) in response to pathogens

• Peripheral blood mononuclear cells (PBMCs): For examining IL1F10's modulatory effects on IL36G-induced responses

• Dendritic cells: For investigating IL1F10's enhancement of LPS-stimulated IL6 production

• B lymphocytes, neutrophils, monocytes: Express IL-1 family receptors and represent physiologically relevant targets

Experimental systems:

• In vitro co-culture systems with mixed immune cell populations provide more physiologically relevant contexts than isolated single-cell type cultures

• Ex vivo tissue explant cultures from relevant disease sites (e.g., psoriatic skin, inflammatory bowel tissues)

• Mouse models with IL1F10 knockout or overexpression for in vivo studies

When designing experiments, researchers should include both positive controls (known IL-1 family cytokines) and negative controls (irrelevant recombinant proteins with similar tags) to account for potential tag-specific effects.

How does the His-tag potentially affect IL1F10 protein function and experimental outcomes?

The presence of a histidine tag on recombinant IL1F10 introduces several methodological considerations:

• Receptor binding: The position of the His-tag (N-terminal versus C-terminal) may differentially impact receptor interactions. C-terminal tags (as in many commercial preparations) generally have minimal interference with IL-36R/IL1RL2 binding, while N-terminal tags might affect the N-terminal regions potentially involved in receptor recognition .

• Protein stability: His-tags may enhance protein solubility and stability in some buffer conditions but might also expose hydrophobic patches leading to aggregation over time.

• Experimental artifacts: His-tags can introduce non-specific binding to metal-containing surfaces or proteins. Control experiments should include:

  • Comparison with tag-cleaved protein when possible

  • Control proteins with identical tags

  • Pre-blocking of potential non-specific binding sites with imidazole in some assays

To rigorously assess potential tag effects, researchers should consider comparing results from multiple versions of the protein (different tag positions or tag-free) when investigating novel IL1F10 functions.

What are the signaling pathways activated or inhibited by IL1F10/IL-38 in different cellular contexts?

IL1F10/IL-38 exhibits context-dependent signaling effects that require comprehensive analysis approaches:

• Receptor interaction: IL1F10 acts as a ligand for IL-36R/IL1RL2, but the downstream signaling differs from other IL-1 family cytokines . Experimental approaches should examine:

  • Receptor binding kinetics using surface plasmon resonance

  • Co-immunoprecipitation of receptor complexes

  • FRET-based approaches to study receptor complex formation

• Downstream pathway modulation:

  • IL1F10 reduces IL22 and IL17A production in T-cells, suggesting inhibition of pathways promoting these cytokines

  • IL1F10 reduces IL36G-induced IL8 production, indicating potential interference with NF-κB or MAP kinase signaling

  • IL1F10 increases IL6 production in dendritic cells after LPS stimulation, suggesting enhancement of specific TLR4-dependent pathways

For rigorous signaling research, phosphoproteomic approaches combined with pathway inhibitors provide the most comprehensive view of IL1F10's complex signaling effects.

What experimental approaches can resolve contradictory findings about IL1F10's anti- versus pro-inflammatory effects?

The apparently contradictory effects of IL1F10 (anti-inflammatory in some contexts, pro-inflammatory in others) require sophisticated experimental approaches:

• Time-course experiments: IL1F10's effects may vary temporally, with initial pro-inflammatory actions followed by resolution phases. Comprehensive time-course studies measuring multiple cytokines simultaneously are essential.

• Cell-specific responses: Different cell types may respond differently to IL1F10. Single-cell analysis approaches including:

  • scRNA-seq of mixed populations after IL1F10 treatment

  • CyTOF analysis of signaling pathway activation in heterogeneous populations

  • Cell type-specific conditional knockout models

• Dose-response considerations: IL1F10 may exhibit hormetic effects (different responses at low versus high concentrations). Full dose-response curves rather than single concentrations should be examined.

• Resolution of inflammation: IL1F10 may promote resolution rather than simply inhibiting inflammation. Specialized pro-resolving mediator (SPM) analysis alongside traditional inflammatory markers provides more comprehensive understanding.

A methodological limitation in many studies is the examination of terminal readouts rather than kinetic parameters. Time-resolved approaches are crucial for understanding IL1F10's complex immunomodulatory profile.

How is IL1F10 expression and function altered in pathological conditions?

IL1F10 has been associated with various pathological conditions that offer important research opportunities:

• Disease associations:

  • Autism Spectrum Disorder

  • Behcet Syndrome

  • Inflammatory conditions where IL-1 family signaling is dysregulated

• Experimental approaches for studying pathological alterations:

  • Quantitative analysis of IL1F10 expression in diseased versus healthy tissues using qPCR and immunohistochemistry

  • Functional assessment of patient-derived IL1F10 versus recombinant protein

  • Genetic association studies examining IL1F10 variants and disease susceptibility

  • Single-cell analysis of IL1F10 expression in complex disease tissues

• Methodological considerations:

  • Control for medication effects in patient samples

  • Appropriate age/sex/genetic background matching of control samples

  • Analysis of both protein and mRNA levels, as discrepancies may indicate post-transcriptional regulation

Single-cell approaches are particularly valuable as IL1F10 may be expressed by specific cellular subsets within complex tissues, and bulk analysis might obscure important disease-associated alterations.

What are the considerations for developing IL1F10-based therapeutic approaches?

Exploring IL1F10 as a therapeutic target requires specific methodological approaches:

• Delivery systems for recombinant IL1F10:

  • PEGylation to extend half-life

  • Nanoparticle encapsulation for targeted delivery

  • Fusion proteins with tissue-targeting domains

• Design of IL1F10 variants with enhanced stability or function:

  • Site-directed mutagenesis to identify and modify key functional residues

  • Development of receptor-selective mutants that retain anti-inflammatory but not pro-inflammatory properties

  • Creation of stabilized versions resistant to proteolytic degradation

• Development of IL1F10-neutralizing approaches for conditions where inhibition is desired:

  • Neutralizing antibodies

  • Receptor antagonists

  • Aptamer-based inhibitors

When developing therapeutic approaches, researchers should consider implementing humanized mouse models expressing human IL1F10 and its receptors to better predict human responses, as species differences in IL-1 family members can be significant.

What are the optimal methods for detecting and quantifying endogenous versus recombinant His-tagged IL1F10 in experimental samples?

Distinguishing endogenous from recombinant His-tagged IL1F10 requires specific methodological considerations:

• Antibody-based detection methods:

  • Anti-IL1F10 antibodies detect both endogenous and recombinant proteins

  • Anti-His tag antibodies specifically detect recombinant protein

  • Dual-color immunofluorescence or western blotting with both antibody types can differentiate populations

• Mass spectrometry approaches:

  • Peptide mass fingerprinting can distinguish recombinant from endogenous proteins

  • Multiple reaction monitoring (MRM) assays can be developed for specific quantification of His-tagged peptides

• ELISA-based detection:

  • Commercial DuoSet ELISA kits are available for human IL1F10/IL-38

  • Modified sandwich ELISA using anti-His capture and anti-IL1F10 detection antibodies specifically measures recombinant protein

A common methodological challenge is cross-reactivity with other IL-1 family members. Validation using IL1F10 knockout samples or recombinant protein standards is essential for ensuring specificity of detection methods.